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Release and Cytokine Production of BmpB from BmpB-Loaded pH-Sensitive and Mucoadhesive Thiolated Eudragit
Abstract:
Swine dysentery is a contagious mucohaemorrhagic colitis of pigs that is caused by anaerobic intestinal spirochaete Brachyspira hyodysenteriae. Recently, an outer membrane lipoprotein of B. hyodysenteriae (BmpB) has been identified, and the mice or pigs immunized with a recombinant BmpB generated antibodies recognizing the native BmpB of B. hyodysenteriae. In this study, we cloned, expressed and purified BmpB protein from E. coli and used it as a vaccine candidate for oral delivery. The BmpB was encapsulated into the pH-sensitive and thiolated Eudragit microspheres (TEMS). The sizes of the microspheres ranged from 5-20 μ. About 22-34% of BmpB were released from the BmpB-loaded TEMS within 24 h at stomach pH 2.0 whereas the release of BmpB from the BmpB-loaded TEMS was 35% in the first one hour and reached 81% within 24 h at intestinal pH 7.2. These data revealed that the BmpB could be protected in the harsh gastric condition. Mucoadhesive experiment in vitro showed that TEMS have high binding affinity with the mucin glycoproteins of porcine intestine. Finally, in vitro production of cytokines from immune cells treated with the BmpB-loaded TEMS suggested that the TEMS would be a promising approach for oral delivery of BmpB as vaccine candidate.
Insights
This study developed a novel oral vaccine delivery system using pH-sensitive microspheres for Brachyspira hyodysenteriae outer membrane lipoprotein BmpB. The system protects the vaccine in the stomach and shows mucoadhesive properties for effective delivery.
Area of Science:
- Veterinary immunology
- Microbiology
- Biomaterials science
Background:
- Swine dysentery is caused by Brachyspira hyodysenteriae.
- Outer membrane lipoprotein BmpB is a potential vaccine candidate.
- Oral delivery systems are needed for improved vaccine efficacy.
Purpose of the Study:
- To develop and evaluate pH-sensitive and thiolated Eudragit microspheres (TEMS) for oral delivery of BmpB.
- To assess the stability and release profile of BmpB from TEMS.
- To investigate the mucoadhesive properties and in vitro immune response of BmpB-loaded TEMS.
Main Methods:
- Cloning, expression, and purification of BmpB protein from E. coli.
- Encapsulation of BmpB into TEMS.
- In vitro release studies at different pH values (2.0 and 7.2).
- In vitro mucoadhesion assay using porcine intestinal mucin.
- In vitro cytokine production assay using immune cells.
Main Results:
- BmpB was successfully encapsulated into TEMS (5-20 μm).
- TEMS protected BmpB in simulated gastric conditions (pH 2.0) with minimal release.
- High BmpB release (81%) occurred at intestinal pH (7.2).
- TEMS demonstrated strong mucoadhesion to porcine intestinal mucin.
- BmpB-loaded TEMS induced cytokine production in vitro, suggesting an immune response.
Conclusions:
- TEMS provide a promising approach for the oral delivery of BmpB as a vaccine candidate against swine dysentery.
- The pH-sensitive and mucoadhesive properties of TEMS enhance oral vaccine delivery.
- Further in vivo studies are warranted to confirm the efficacy of this oral vaccine system.
